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배양대뇌신경세포 저산소증모델에서 반하여 의한 유전자표현의 변화

Microarray Analysis of Gene Expression Affected by Water-extracts of Pinelliae rhizoma in a Hypoxic Model of Cultured Rat Cortical Cells

초록

본 연구는 저산소증에서 반하가 대뇌신경세포의 유전자 표현에 미치는 영향을 알아보기 위하여 배양한 $E_{18}$의 흰쥐 대뇌 신경세포를 반하로 처리하고, 저산소증을 유도한 후 microarray 기법으로 유전자 표현 변화를 조사하였다. Microarray 결과 tubb5, tgfa, ptpn11, n-ras, pdgfa 등 세포의 성장 분화에 관여하는 유전자들의 표현이 증가하였으며, 세포 자연사를 억제하는 mcl-1 유전자의 표현 또한 증가하였다. 한편 세포 자연사를 유도하는 tieg 유전자는 표현이 감소하였다(Fig. 3). 반하에 의하여 수많은 유전자의 표현이 변화되었고, 세포사를 촉진하는 유전자의 표현이 크게 증가되는 경우(예, alox12, faf1)도 있어 본 연구결과만으로 일반적인 결론을 유도하기는 어려웠다. 그러나 대략적으로 반하는 저산소증에서 주로 세포의 성장과 분화를 유지하고, 세포 자연사를 방지하는 유전자들의 표현을 증가시켜 신경 세포사를 보호하는 것으로 이해된다.

Abstract

Pinelliae rhizoma (Pr, 半夏) is a traditional medicine used in the treatment of incipient stroke. We investigated the effects of Pr on gene expression in a hypoxic model using cultured rat cortical cells. Pr (2.5 $\mu$g/ml) was added to the culture medium on DIV 12. A hypoxic shock (2% 0$_2$/5% CO$_2$, 37$^{\circ}$C, 3 hr) was given two days later (on DIV 14), and total mRNAs were isolated at 24 hr post-shock from both Pr-treated samples and untreated control cultures. Microarray using TwinChip $^{TM}$ Rat-5K (Digital Genomics, Seoul) indicated that Pr upregulated genes for cell growth and differentiation (tubb5, tgfa, ptpn11, n-ras, pdgfa) and antiapoptosis (mcl-1), while downregulating the apoptosis-induced gene (tieg). Therefore, it is interpreted that Pr protects neurons from hypxoic shock by maintaining cell growth and differentiation and by preventing apoptosis.

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